【摘要】 背景:已证实在特定条件下,脐血干细胞经刺激可诱导分化为神经干细胞,用于移植治疗中枢神经系统疾病具有明显优越性。目的:观察脐血干细胞移植后受者血液中神经元特异性烯醇化酶和β2-微球蛋白的表达变化。设计:病例分析。对象:2007-03/07深圳市第六人民医院康复医学科行脐血干细胞移植的54例神经系统疾病患者,对治疗及实验均知情同意。脐血干细胞由深圳北科生物技术有限公司提供。方法:每例患者用2mL注射器向蛛网膜下腔推注脐血干细胞悬液1.0～2.0mL,细胞数(3.0～4.0)×1010L-1,每5d移植1次,4次为1个疗程。每例患者取左侧屈膝位,定位L3/4或L2/3椎间隙,麻醉后由椎间隙进硬膜外腔,再继续进针至有落空感时抽出针芯,有清亮脑脊液流出,留取脑脊液标本送检。每例患者静脉采血3.0～4.0mL/次,离心后分离血清,-20℃保存,避免反复冻融。主要观察指标:ELISA法和免疫组化法测定神经元特异性烯醇化酶的表达,RIA法测定β2-微球蛋白的表达。结果:与移植前比较,移植后血清和脑脊液中神经元特异性烯醇化酶质量浓度均无明显变化(P>0.05);移植后血清中β2-微球蛋白质量浓度无明显变化(P>0.05),脑脊液中β2-微球蛋白质量浓度显著升高(P<0.01)。移植前后脑脊液沉淀细胞神经元特异性烯醇化酶免疫组化显色差异有明显变化(P<0.05)。结论:脐血干细胞移植对脑脊液和血清中的神经元特异性烯醇化酶表达无明显影响,脑脊液沉淀细胞可能增殖分化为神经元样细胞。移植后β2-微球蛋白在正常范围内的升高,从临床角度说明脐血干细胞免疫排斥反应小及应用的安全性。更多还原

【Abstract】 BACKGROUND:Under special conditions,umbilical cord blood stem cells can differentiate into neural stem cells(NSCs),and has significant superiority on treatment of central nervous system disease.OBJECTIVE:To investigate the changes in expression of neuron-specific enolase(NSE) and β2-microglobulin(β2-MG) in recipient blood following umbilical cord blood stem cell transplantation.DESIGN:Case analysis.PARTICIPANTS:A total of 54 nervous system disease patients undergoing umbilical cord blood stem cell transplantation were recruited at the Department of Rehabilitation Medicine,Shenzhen Sixth People’s Hospital from March to July 2007.The patients signed informed consents.Umbilical cord blood stem cells were supplied by Shenzhen Beike Biotechnology Co.,Ltd.METHODS:1.0-2.0 mL of umbilical cord blood stem cells was infused in the subarachnoid cavity using 2 mL syringe in each patient.Cell number was(3.0-4.0)×1010/L,once every 5 days,4 times as a course.At left genuflex position and L3/4 or L2/3 intervertebral space in each patient,the needle entered following anesthesia from the intervertebral space into the epidural cavity,and needle core was extracted when feels nothing.Clear cerebrospinal fluid was effluent and collected for further use.3.0-4.0 mL/times of venous blood was harvested in each patient,centrifuged,stored at-20 ℃ following serum isolation to avoid repeatedly freeze thawing.MAIN OUTCOME MEASURES:Expression of NSE was determined by using ELISA and immunohistochemistry.β2-MG expression was determined by RIA.RESULTS:There were no significant changes in NSE mass concentration in serum and cerebrospinal fluid following transplantation(P > 0.05).No significant difference in β2-MG mass concentration was determined following transplantation in serum(P > 0.05).β2-MG mass concentration was significantly increased in the cerebrospinal fluid(P < 0.01).Before and after transplantation,cerebrospinal fluid precipitation cell of the neuron-specific enolase immunohistochemical color difference was significantly changed(P < 0.05).CONCLUSION:Umbilical cord blood stem cell transplantation has no effects on NSE expression in cerebrospinal fluid and serum.Cerebrospinal fluid sediment cells can proliferate and differentiate into neuron-like cells.Increased β2-MG expression(in normal range) indicated that the small rejection and security application of umbilical cord blood stem cells in clinic.更多还原